The present application is a 371 application of PCT/JP00/02492 filed Apr. 17, 2000.
The present invention relates to a novel protein having amidase activity that stereoselectively hydrolyzes xcex1-amino acid amides and xcex1-hydroxy acid amides, and a gene encoding said protein.
It is known that optically active xcex1-amino acids and xcex1-hydroxy acids can be produced using microorganism or enzyme threof as a catalyst to hydrolyze xcex1-amino acid amides and xcex1-hydroxy acid amides. Examples of optically active xcex1-amino acids are described in Japanese Patent Application Laying-Open (kokai) Nos. 61-293394 and 62-55097; optically active xcex1-hydroxy acids are described in Japanese Patent Application Laying-Open (kokai) No. 2-84198.
Amidase genes cloned via gene recombination techniques can greatly enhance the catalytic capability of microorganisms compared with the conventional methods, because it enables the existence of multiple copies of amidase genes in a cell.
In order to prepare a microbial catalyst having higher catalytic activity, we have completed the present invention by cloning an amidase gene which stereoselectively hydrolyzes xcex1-amino acid amides and xcex1-hydroxy acid amides.
The present invention provides the followings:
(1) A protein defined in the following (a) or (b):
(a) a protein having an amino acid sequence of SEQ ID NO: 2;
(b) a protein having an amino acid sequence that differs from the amino acid sequence of SEQ ID NO: 2 by deletion, replacement or addition of one or more amino acids, and having amidase activity that stereoselectively hydrolyzes xcex1-amino acid amides and xcex1-hydroxy acid amides.
(2) A gene which encodes the protein of (1)
(3) The gene of (2) which having a nucleotide sequence of nucleotides 361 to 1305 of SEQ ID NO: 1.
(4) A gene capable of hybridizing to the nucleotide sequence of nucleotides 365 to 1305 of SEQ ID NO: 1 or fragments thereof, and encodes a protein having amidase activity that stereoselectively hydrolyzes xcex1-amino acid amides and xcex1-hydroxy acid amides.
(5) A recombinant vector to which the gene of any one of (2) to (4) is linked.
(6) A transformant which is prepared by introducing the recombinant vector of (5) into a host microorganism.
(7) A method for producing a protein having amidase activity which comprises a culturing process of the transformant of (6)
When a protein having amidase activity, which has been obtained by culturing the above transformant, acts on xcex1-amino acid amides, optically active xcex1-amino acids corresponding thereto can be produced; and when the same protein acts on xcex1-hydroxy acid amides, optically active xcex1-hydroxy acids corresponding thereto can be produced.
This specification includes part or all of the contents disclosed in the specification and/or drawings of Japanese Patent Application No. 11-109328, which is a priority document of the present application.